The CCR Confocal Microscopy Core provides new "dimensions" in imaging

Thursday, October 27, 2011 — Core Poster Session
10:00 a.m. – Noon	South Lobby of Building 10	NCI	CORE-9

Authors

• S Garfield
• P Mannan
• L Lim

Abstract

The Confocal Core provides state-of-the-art microscopic analyses to better understand critical biological structures and cellular processes involved in cancer. Confocal Microscopy is a valuable research tool for imaging fluorescently labeled specimens, permitting accurate, non-invasive optical sectioning. Techniques used in this facility are: 1) co-localization of fluorescent fusion proteins with organelles, 2) demonstration of membrane ruffling, cytoskeletal organization, focal adhesions and other cell morphology, 3) live time-lapse translocation of fluorescent fusion proteins, 4) fluorescent indicators of oxidative stress in live cells, 5) 4D imaging of cell division and 6) Second Harmonic Generation imaging (SHG) of whole live tissue/organ. Confocal systems are suitable for Fluorescent Recovery After Photobleaching (FRAP) and Fluorescent Resonance Energy Transfer (FRET) experiments. The Zeiss LSM 510 NLO with Meta detector, for separating fluorophores with close emission spectra, was added in 2004. This 2-photon system images thick tissue with minimum tissue damage with an x,y scanning stage and environmental chamber. The new Zeiss 710 with 2-photon laser provides 1p and 2p Fluorescence Lifetime Imaging (FLIM), has x,y scanning stage for tiling, environmental chamber plus non-descanned detectors (NDDs) for deeper imaging, and 32 channel PMT. Specialized software and computers are available to users.

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